电针对脊髓损伤星形胶质细胞增生及其NGF表达的影响

目的研究脊髓损伤后电针治疗对星形胶质细胞增生及其内源性神经生长因子(nerve growth factor,NGF)表达的影响.方法选用成年雌性Wistar大鼠,随机分为3组.A组为正常对照组,B组、C组为下胸段脊髓不完全损伤.B组损伤后不治疗,C组损伤后给予督脉电针治疗.损伤后3 d、1 、2或4周应用免疫组化染色分别观察损伤脊髓胶质原纤维酸性蛋白(glial fibroblast acid protein,GFAP)和NGF表达的变化.结果 B组术后3 d,GFAP阳性细胞明显增多, 2周后开始减少,4周时仍有较多的阳性细胞;C组GFAP阳性细胞明显少于B组,1周时达高峰.脊髓损伤后NGF表达呈逐渐增加的趋势.C组NGF的表达明显高于B组,且一直保持在较高水平.NGF阳性细胞大部分与GFAP阳性细胞形态相似.结论电针治疗能减少星形胶质细胞增生,促进内源性NGF的合成,从而创造了有利于神经再生的微环境.     

PDAPP转基因小鼠脑组织内反应性星形胶质细胞的活化程度明显升高

目的　研究PDAPP转基因小鼠脑组织内反应性星形胶质细胞的活化程度。方法　通过免疫组织化学染色方法检测小鼠脑组织内反应性星形胶质细胞表达胶质纤维酸性蛋白 (GFAP)的情况 ,比较PDAPP转基因小鼠和C5 7 BL非转基因小鼠脑组织反应性星形胶质细胞的活化程度。结果　PDAPP转基因小鼠脑组织内反应性星形胶质细胞表达GFAP的水平明显高于C5 7 BL非转基因小鼠。结论　PDAPP转基因小鼠脑组织内存在明显的神经炎症反应     

LPS诱导海马星形胶质细胞活化与活化的星形胶质细胞对海马神经元的影响

探讨脂多糖(Lipopolysaccharide,LPS)对长时间存活大鼠海马内星形胶质细胞的反应以及对神经元的影响。方法:本实验用10只健康成年雄性SD大鼠,海马CA3区注射LPS 10μ1．7和14d后,尼氏染色观察神经元的变化,免疫组织化学染色结合图像分析方法观察海马CA3区注射部位胶质纤维酸性蛋白(glial fibrillary acidic protein GFAP)、的表达变化。结果:脂多糖可促进海马星形胶质细胞的活化,但并不能引起海马区神经元的损伤。结论:星形胶质细胞在脑损伤后的脑内炎症反应起了一定的作用,但并不能引起神经元的损伤。     

NDRG2与GFAP在不同脑区星形胶质细胞的表达与分布

目的:观察NDRG2(N-myc下游调节基因2)与GFAP(胶质纤维酸性蛋白)在不同脑区星形胶质细胞的表达与分布。方法:利用免疫荧光NDRG2与GFAP双标技术以及Western Blot技术观察皮层、海马及纹状体等不同脑区星形胶质细胞NDRG2和GFAP的表达与分布。结果:免疫荧光结果显示NDRG2阳性细胞广泛而均匀地分布于不同脑区,并与GFAP存在较好的共定位;NDRG2与GFAP标记的星形胶质细胞形态不尽相同。Western Blot结果显示NDRG2在皮层中表达比海马和纹状体多,而GFAP在海马中表达比皮层和纹状体多。结论:NDRG2广泛表达于不同脑区星形胶质细胞,并于GFAP存在较好的共定位。     

正加速度暴露对大鼠海马星形胶质细胞GFAP表达的影响

目的探讨正加速度( Gz)重复暴露后不同时间海马星形胶质细胞GFAP表达的变化.方法 SD大鼠60只,随机分成对照组、 Gz重复暴露后1h、6h、12h、24h和48h组,每组10只.采用动物离心机,建立 Gz引发急性脑缺血模型;应用免疫组织化学技术,分别检测 Gz重复暴露后不同时间,海马星形胶质细胞GFAP的表达状况.结果海马星形胶质细胞GFAP阳性细胞数,在 Gz暴露后1h即显著增加,于12h达到高峰,而后逐渐下降,48h仍维持在较高水平,实验组与对照组比较,有显著性差异.结论 Gz重复暴露导致海马星形胶质细胞GFAP表达上调,可能对神经元的缺血损伤起保护作用.     

X射线对体外培养星形胶质细胞增生活化和分泌的影响

目的观察X射线照射对培养星形胶质细胞(astrocyte,AS)增殖和分泌的影响。方法建立体外培养大鼠纯化的AS物理损伤模型(划痕损伤模型),分为对照组、划痕组和放疗组。利用免疫荧光观察胶质细胞胶质纤维酸性蛋白(glial fibrillary acidic protein,GFAP)和5-溴脱氧尿嘧啶核苷(5-bromodeoxy uridine,BrdU)的表达,利用RT-PCR观察细胞因子IL-6和TNF-a的表达。结果划痕损伤刺激能使培养AS反应性增生活化。损伤后6h出现IL-6、TNF-a表达水平增高,24h后损伤周围BrdU阳性细胞率明显增加。通过X射线照射能抑制AS的BrdU表达,但并不能抑制损伤后IL-6和TNF-a的表达。结论X射线照射可以通过调控细胞周期,有效抑制损伤后AS的反应性增生,但不能对活化AS的IL-6和TNF-a的表达起到抑制作用。     

异丙酚对局灶性脑缺血／再灌注星形胶质细胞GFAP表达的影响

目的：探讨异丙酚对局灶性脑缺血／再灌注后星形胶质细胞胶质纤维酸性蛋白（GFAP）表达的影响。方法：大脑中动脉插线法制作大鼠局灶性脑缺血／再灌注模型。观察脑缺血2h再灌注24h后神经功能损害改变并评分,并采用免疫荧光组织化学法检测大鼠齿状回GFAP蛋白的表达。结果：缺血／再灌注后可诱导大鼠齿状回GFAP表达明显增强,异丙酚可抑制缺血／再灌注后GFAP的表达,明显改善大鼠神经功能损害（P〈0．05或0.01）。结论：异丙酚通过抑制脑缺血后星形胶质细胞GFAP的过度表达发挥抗脑缺血损伤保护神经元作用。     

胃伤害性刺激诱导大鼠脑干星形胶质细胞GFAP蛋白表达及其与神经元的关系

研究大鼠在福尔马林诱发胃伤害性刺激时脑干内星形胶质细胞及神经元的变化。应用免疫组织化学三重标记法在脑原位切片同时显示脑干内Fos蛋白,胶质原纤维酸性蛋白（GFAP）,酪氨酸羟化酶（TH）的表达,结果显示：1、在福尔马林诱发胃伤害性刺激后,脑干胶质细胞GFAP表达阳性,并表现出明显的核团或亚核定位特点,在延髓内脏带（MVZ0,中缝大核（RMg),蓝斑（LC）,臂旁外侧核（LPB）,中缝背核（DR）,中脑导水管周围灰质腹外侧区（vlPAG),上丘中灰层（IngSC)等脑区有较多的Fos阳性细胞,而且Fos阳性表达的分布与上述GFAP阳性分布基本一致;2、MVZ,LC,DR,vlPAG等部位有大量Fos及TH双标阳性神经元,周围有密集的GFAP阳性细胞;3、随着刺激后存活时间的变化,GFAP与Fos阳性细胞的反应均经历逐渐升高后又渐降低直至消失的变化。结果表明：上述核团的神经元和星形胶质细胞可能同时参与了内脏痛及其调节过程。     

猫视皮层星形胶质细胞的老年性变化

电生理研究结果显示,在衰老过程中猫的视皮层神经元对视觉刺激的反应性出现显著的功能衰退,是否这种功能性衰退伴随胶质细胞活动的改变尚无直接的实验证据。以前期电生理实验猫为材料,用免疫形态学方法比较青年猫和老年猫初级视皮层内星形胶质细胞的活动状况。利用Nissl染色显示猫初级视皮层组织结构,用免疫组织化学方法（SABC法）显示GFAP免疫阳性（GFAP-IR）星形胶质细胞。光镜下观察、拍照,对GFAP-IR细胞计数并换算成密度,测量GFAP-IR直径取平均值。老年猫初级视皮层灰质各层及白质内的GFAP-IR细胞密度比青年猫的显著升高（p〈0.001）。与青年猫相比,老年猫视皮层灰质和白质中GFAP-IR细胞的平均直径均比青年猫的显著增大（p〈0.0001）,且老年猫视皮层内GFAP阳性免疫反应较青年猫的明显增强。老年猫初级视皮层神经元功能衰退伴随着星形胶质细胞活动的增强,胶质细胞活动增强有助于神经元之间的信息交流,因而可能对衰老过程中神经元的功能衰退起补偿作用。     

Olomoucine对星形胶质细胞增殖及活化分泌的影响

目的观察细胞周期抑制剂olomoucine对培养星形胶质细胞机械损伤后增殖和活化分泌的影响。方法建立体外培养大鼠纯化的AS物理损伤模型（划痕损伤模型）,分为对照组、划痕组和olomoucine干预组。利用免疫荧光细胞化学方法观察损伤后GFAP表达;利用RT-PCR观察细胞因子IL-6和TNF-α的表达情况。结果划痕损伤刺激能使培养AS反应性增生活化。损伤早期出现IL-6、TNF-α表达水平增高,从损伤后12h开始出现损伤边缘区AS胞体肥大,数目增加;给予olomoucine干预后,细胞数目减少、体积明显减小,损伤后IL-6和TNF-α表达也显著下降。结论损伤刺激可促使AS活化,并发生反应性胶质增生;CDK选择性细胞周期抑制剂olomoucine可以通过调控细胞周期,有效抑制损伤后星形胶质细胞的反应性增生,并能对AS的活化起到抑制作用。     

Synergistic Cooperation between Methamphetamine and HIV-1 gsp120 through the P13K/Akt Pathway Induces IL-6 but not IL-8 Expression in Astrocytes

HIV-1 envelope protein gp120 has been extensively studied for neurotoxic effects that have been attributed to the increased expression of various proinflammatory cytokines in the CNS. Recently we have shown that methamphetamine (MA) also increases expression of proinflammatory cytokines in astrocytes. However, combined effect of gp120 and MA is not known. The present study was undertaken to determine cumulative effect and the mechanism(s)/pathways involved in the functional interaction between gp120 and MA in SVGA astrocytes. Our results clearly suggest that gp120 and MA affect IL-6 but not IL-8 in a synergistic manner and this synergy was mediated by PI3K/Akt and NF-κB pathways. Inhibition of either of these pathways could abrogate the increased expression of IL-6 due to MA or gp120 alone, as well as the increased expression of IL-6 when the astrocytes were treated with both gp120 and MA. These results were confirmed by both, using chemical inhibitors/siRNA as well as western blotting. This study therefore provides novel information regarding the interaction between MA and gp120 in terms of the expression of IL-6 and the mechanisms underlying potential synergy between MA and gp120 in astrocytes.     

Serum NGF,BDNF and IL-6 Levels in Postpartum Mothers As Predictors of Infant Development: The Influence of Affective Disorders

Background

Early adverse experiences are associated with increased risk of developing psychiatric disorders, although little is known about the neurobiological mediators involved. The mechanisms by which early environmental influences may mediate vulnerability in the development of offspring await further investigation. The present study correlated the NGF, BDNF, IL-6 and cortisol levels of mothers with postpartum affective disorders (PPAD) with infant development.

Methods

A longitudinal study was performed with 152 pregnant women and their infants. Between 60 and 120 days after delivery, women were interviewed and provided biological samples for biochemical analysis, and the infants were examined for neurobiological-motor development.

Results

Overall, the mothers'' history of affective disorders, PPAD and anxiety disorder were associated with infant motor development. Using an adjusted linear regression analysis, PPAD (p = 0.049), maternal anxiety disorder (p = 0.043), NGF level (p = 0.034) and infant cortisol level (p = 0.013) were associated with infant motor development. Using a factorial analysis of primary components, two components were retained. The psychological factor was characterized by a positive loading of a history of affective disorder, PPAD and anxiety disorder. For the biological factor, infant cortisol adhered negatively with infant motor development, but NGF was positively associated. The psychological factor had a negative association, but the biological factor had a positive association with infant motor development.

Conclusions

There are few studies that have focused on the relationship of biomarkers and infant neurodevelopment. Our study points that psychological and biological factors are associated with infant motor development, however the causal relationship between these factors is still to be defined.     

IL-1，IL-6，IL-8 与胃癌的关系

胃癌是常见的恶性肿瘤之一,在我国其发病率居各类肿瘤前列,导致的死亡人数占所有肿瘤的四分之一,而且每年有近40万新增的胃癌病人,但是其早期诊断率低于20%,胃癌已经成为危害人民健康的最严重的疾病之一。白细胞介素(interleukin,IL)作为在白细胞或免疫细胞间相互作用的淋巴因子,不仅在介导T、B细胞活化、增殖与分化以及炎症反应中起着重要作用,近年来,越来越多的学者发现其与肿瘤的发生及发展也有着密不可分的联系。目前为止,已经发现了29种白细胞介素,分别被命名为IL-1~IL-29,它们各自承担着相应的使命。国内外大量实验及文献表明,IL-1,IL-6,IL-8和胃癌有着密切的关系,这对于胃癌的早期诊断及治疗提出了新的思路,进而提高胃癌的早期诊断率,改善胃癌的治疗状况。本文对IL-1,IL-6,IL-8的来源、分子结构及受体方面进行简要概述,同时阐述了其生物学特性及与胃癌的关系。     

血清白介素-6和白介素-10浓度的变化与冠心病

目的探讨白介素-6(IL-6)、白介素-10(IL-10)在冠心病心绞痛患者血中的变化规律。方法检测25例不稳定型心绞痛、23例稳定型心绞痛患者及22例正常对照者组血中IL6、IL-10浓度并进行比较。结果不稳定型心绞痛组、稳定型心绞痛组及正常对照组血中IL-6分别为(298.6±52.4)、(143.2±46.9)、(75.1±32.7)pg/m l;不稳定型心绞痛组分别高于稳定型心绞痛组及正常对照组,差异均有非常显著性(均为P<0.001)。不稳定型心绞痛组、稳定型心绞痛组及正常对照组血IL-10分别为(173.7±30.9)、(80.4±15.6)、(38.2±7.5)pg/m l,不稳定型心绞痛组分别高于稳定型心绞痛组及正常对照组(P<0.01)。结论冠心病患者血清IL-6、IL-10的浓度升高。     

Expression and structure of the human NGF receptor

The nucleotide sequence for the human nerve growth factor (NGF) receptor has been determined. The 3.8 kb receptor mRNA encodes a 427 amino acid protein containing a 28 amino acid signal peptide, an extracellular domain containing four 40 amino acid repeats with six cysteine residues at conserved positions followed by a serine/threonine-rich region, a single transmembrane domain, and a 155 amino acid cytoplasmic domain. The sequence of the extracellular domain of the NGF receptor predicts a highly ordered structure containing a negatively charged region that may serve as the ligand-binding site. This domain is conserved through evolution. Transfection of a full-length cDNA in mouse fibroblasts results in stable expression of NGF receptors that are recognized by monoclonal antibodies to the human NGF receptor and that bind [125I]NGF.     

Clinical Relevance of VPAC1 Receptor Expression in Early Arthritis: Association with IL-6 and Disease Activity

Background

The vasoactive intestinal peptide (VIP) receptors VPAC1 and VPAC2 mediate anti-inflammatory and immunoregulatory responses in rheumatoid arthritis (RA). Data on the expression of these receptors could complement clinical assessment in the management of RA. Our goal was to investigate the correlation between expression of both receptors and the 28-Joint Disease Activity Score (DAS28) in peripheral blood mononuclear cells (PBMCs) from patients with early arthritis (EA). We also measured expression of IL-6 to evaluate the association between VIP receptors and systemic inflammation.

Methods

We analyzed 250 blood samples collected at any of the 5 scheduled follow-up visits from 125 patients enrolled in the Princesa Early Arthritis Register Longitudinal study. Samples from 22 healthy donors were also analyzed. Sociodemographic, clinical, and therapeutic data were systematically recorded. mRNA expression levels were determined using real-time PCR. Then, longitudinal multivariate analyses were performed.

Results

PBMCs from EA patients showed significantly higher expression of VPAC2 receptors at baseline compared to healthy donors (p<0.001). With time, however, VPAC2 expression tended to be significantly lower while VPAC1 receptor expression increased in correlation with a reduction in DAS28 index. Our results reveal that more severe inflammation, based on high levels of IL-6, is associated with lower expression of VPAC1 (p<0.001) and conversely with increased expression of VPAC2 (p<0.001). A major finding of this study is that expression of VPAC1 is lower in patients with increased disease activity (p = 0.001), thus making it possible to differentiate between patients with various degrees of clinical disease activity.

Conclusion

Patients with more severe inflammation and higher disease activity show lower levels of VPAC1 expression, which is associated with patient-reported impairment. Therefore, VPAC1 is a biological marker in EA.     

重组人白细胞介素－6与肿瘤坏死因子突变体融合蛋白的构建及表达

针对肿瘤坏死因子（ＴＮＦ）在肿瘤治疗剂量下产生的严重毒副作用及一些肿瘤细胞上白细胞介素－６（ＩＬ－６）受体明显增高的事实,根据ＴＮＦ结构与功能研究的最新信息,利用ＰＣＲ技术,对人ＴＮＦα基因进行了改造,并将其与人ＩＬ－６成熟肽编码区ｃＤＮＡ通过人工接头进行融合。融合蛋白在大肠杆菌中表达后,Ｗｅｓｔｅｒｎｂｌｏｔ分析表明,分子量约为３７ｋＤ;活性检测结果证实,该融合蛋白兼具有ＴＮＦ抗肿瘤活性和结合ＩＬ－６受体的能力,在高表达ＩＬ－６受体的人骨髓瘤细胞上测得的细胞毒活性较同样位点突变的ＴＮＦ高约３倍。     

内源性IL-17诱导MIP-2与IL-6表达在衣原体肺炎中促进中性粒细胞循环

目的 探讨衣原体肺炎中白细胞介素-17（interleukin -17,IL-17）对中性粒细胞（polymorphonuclear leucocyte,PMN）循环的调节作用及机制.方法 用40 μl含1×10^3包涵体形成单位（inclusion-forming units,IFU）的衣原体鼠肺炎株（Chlamydia muridarum,Cm）呼吸道感染BALB/c小鼠,诱导鼠衣原体肺炎.用抗鼠IL-17单克隆抗体吸入中和内源性IL-17,以相应独特型抗体（IgG2α）作为对照.用RT-PCR检测小鼠肺组织及肺上皮细胞系巨噬细胞炎性蛋白-2 （macrophage inflammatory protein-2,MIP-2）和IL-6 mRNA的表达.取小鼠支气管肺泡灌洗液细胞染色计数PMN,感染肺组织进行病理染色.结果 衣原体肺炎中,内源性IL-17中和小鼠肺组织PMN浸润显著降低,支气管肺泡灌洗液PMN数量显著低于对照组.IL-17与TNF-α协同可上调肺上皮细胞MIP-2和IL-6 mRNA表达,且内源性IL-17中和小鼠肺组织MIP-2和IL-6表达显著降低.结论 衣原体肺炎中IL-17通过促进肺组织细胞分泌趋化性细胞因子MIP-2和前症性细胞因子IL-6,诱导PMN循环,参与宿主抗衣原体炎性应答.     

Expression of anti human IL-4 and IL-6 scFvs in transgenic tobacco plants

The two murine single-chain Fv (scFv) genes against human interleukin IL-4 and IL-6 cytokines were cloned in a plant expression vector (pGEJAE1) and mobilized to Agrobacterium tumefaciens. Tobacco leaf discs were co-cultured with Agrobacterium and transferred to selective media for regeneration. The tobacco in vitro plants produced scFvs against human IL-4 and IL-6. Only 8% of transformed plants expressing anti-IL-4 scFv were obtained versus 76% of transformed plants expressing anti-IL-6 scFv. In addition, some plants producing anti-IL-4 and anti-IL-6 scFvs aged more rapidly in in vitro conditions and in greenhouse pots than did control plants. Western blot analysis showed that the transformed Nicotiana tabacum plants contained proteins with an apparent molecular mass on electrophoresis of ca. 32 kDa, corresponding to the predicted size of the scFvs. As entire plant root seemed to accumulate more scFv than did leaves, we decided to continue working with isolated roots. Anti-IL-6 scFvs were detected in cultivated roots and their culture media. Functional anti-IL-6 scFv accounted for 0.16–0.18% of total soluble proteins. The affinity of the anti-IL-6 scFv produced in plants and measured by Biacore was similar to that of scFv produced in Escherichia coli. The high levels of antibody accumulation in isolated roots and secretion into the medium demonstrate the potential for producing recombinant protein in bioreactor systems.these authors contributed equally to this workthese authors contributed equally to this work